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谷氨酰胺(glutamine,Gln)在细胞生长和代谢过程中起重要作用,能够激活磷脂酰肌醇-3-激酶(phosphatidylinositol-3-kinase,PI3K)/蛋白激酶B(protein kinase B,Akt)/哺乳动物雷帕霉素蛋白(mammalian target of rapamycin,m TOR)信号通路。多种病毒能通过PI3K/Akt信号通路维持细胞生存和抑制细胞凋亡,维持病毒的感染。为了探讨Gln在痘苗病毒(vaccinia virus strain western reserve,WR)感染人肺癌细胞A549过程中的调控作用及其潜在机制,该文通过蛋白免疫印迹(Western blot)检测PI3K/Akt信号通路蛋白质磷酸化水平,流式细胞术检测细胞阳性率,结晶紫染色检测WR滴度,实时定量PCR(quantitative Real-time PCR,q PCR)检测WR基因E3L和A46R m RNA水平。结果显示,Gln处理后PI3K/Akt信号通路蛋白质磷酸化水平显著升高。抑制PI3K/Akt信号通路后,细胞阳性率显著降低(P<0.05),WR滴度显著降低(P<0.05),WR基因E3L和A46R m RNA水平显著降低(P<0.05)。该研究结果表明,Gln通过激活PI3K/Akt信号通路从而促进A549细胞中WR的复制。
Glutamine (Gln) plays an important role in the process of cell growth and metabolism, and can activate the phosphorylation of PI3K / Akt / Mammalian mammalian target of rapamycin (mTOR) signaling pathway. A variety of viruses through the PI3K / Akt signaling pathway to maintain cell survival and inhibition of apoptosis, to maintain the virus infection. In order to investigate the role of Gln in the pathogenesis of human lung cancer A549 cells infected by vaccinia virus strain western reserve (WR), the phosphoproteome , The positive rate of cells was detected by flow cytometry, the WR titer was detected by crystal violet staining, and the levels of E3L and A46R m RNA of WR gene were detected by quantitative real-time PCR (qPCR). The results showed that phosphorylation of PI3K / Akt signaling pathway was significantly increased after Gln treatment. The inhibitory effect of PI3K / Akt signaling pathway was significantly decreased (P <0.05), the WR titer was significantly decreased (P <0.05), and the levels of WR gene E3L and A46R m RNA were significantly decreased (P <0.05). The results suggest that Gln promotes WR replication in A549 cells by activating the PI3K / Akt signaling pathway.